Circular RNA AFF4 modulates osteogenic differentiation in BM-MSCs by activating SMAD1/5 pathway through miR-135a-5p/FNDC5/Irisin axis.

Bone marrow-derived mesenchymal stem cells (BM-MSCs), the common progenitor cells of adipocytes and osteoblasts, have been recognized as the key mediator during bone formation. Herein, our study aim to investigate molecular mechanisms underlying circular RNA (circRNA) AFF4 (circ_AFF4)-regulated BM-MSCs osteogenesis. BM-MSCs were characterized by FACS, ARS, and ALP staining. Expression patterns of circ_AFF4, miR-135a-5p, FNDC5/Irisin, SMAD1/5, and osteogenesis markers, including ALP, BMP4, RUNX2, Spp1, and Colla1 were detected by qRT-PCR, western blot, or immunofluorescence staining, respectively. Interactions between circ_AFF4 and miR-135a-5p, FNDC5, and miR-135a-5p were analyzed using web tools including TargetScan, miRanda, and miRDB, and further confirmed by luciferase reporter assay and RNA pull-down. Complex formation between Irisin and Integrin αV was verified by Co-immunoprecipitation. To further verify the functional role of circ_AFF4 in vivo during bone formation, we conducted animal experiments harboring circ_AFF4 knockdown, and born samples were evaluated by immunohistochemistry, hematoxylin and eosin, and Masson staining. Circ_AFF4 was upregulated upon osteogenic differentiation induction in BM-MSCs, and miR-135a-5p expression declined as differentiation proceeds. Circ_AFF4 knockdown significantly inhibited osteogenesis potential in BM-MSCs. Circ_AFF4 stimulated FNDC5/Irisin expression through complementary binding to its downstream target molecule miR-135a-5p. Irisin formed an intermolecular complex with Integrin αV and activated the SMAD1/5 pathway during osteogenic differentiation. Our work revealed that circ_AFF4, acting as a sponge of miR-135a-5p, triggers the promotion of FNDC5/Irisin via activating the SMAD1/5 pathway to induce osteogenic differentiation in BM-MSCs. These findings gained a deeper insight into the circRNA-miRNA regulatory system in the bone marrow microenvironment and may improve our understanding of bone formation-related diseases at physiological and pathological levels.

[Analysis of related factors of deep venous thrombosis after spinal cord injury].

OBJECTIVE: To explore the high-risk factors of deep vein thrombosis (DVT) formation in patients after spinal cord injury (SCI) and to provide some reference value for the prevention of DVT. METHODS: Eighty-five patients with spinal cord injury caused by thoracolumbar burst fracture from January 2016 to December 2017 were selected as subjects. All patients were followed up for 6 months, of which 5 cases were lost and 80 cases were finally included. According to whether there was deep vein thrombosis, the patients were divided into DVT group (35 cases) and control group (45 cases). The clinical data were statistically analyzed by SPSS 22.0 statistical software, and the high-risk factors of spinal cord injury were analyzed by multivariate Logistic regression. RESULTS: Thirty-five of the 80 patients with spinal cord injury developed deep venous thrombosis (incidence rate was about 43.7%). The average age of DVT group [(47.77±10.76) years ] was higher than that of non-DVT group [(37.35±10.20) years ], and there was a significant difference between two groups (χ2=19.56, P<0.000). Patients with body mass index≥28 kg/m2, combined with diabetes mellitus, smoking history, no limb pressure therapy and ankle pump exercise had a high incidence of DVT. The incidence of ASIA grade A in the spinal cord injury was higher in the DVT group (48.5%) than in the non-DVT group (20%) (P<0.05). The ROC curve was drawn by MedCalc 15.8 software : when the age was more than 45 years old, the incidence of deep venous thrombosis was higher, the specificity was 84.4%, and the sensitivity was 72.6%. Multivariate Logistic regression analysis showed the following factors : patient age >45 years [OR=1.665, 95% CI (1.102-2.516)], diabetes history [OR=3.273, 95% CI (1.291-8.295)], smoking history [OR=3.302, 95% CI (1.267-8.605)], spinal cord injury ASIA grade A [OR=5.736, 95% CI (3.152-11.74)], non-limb air pressure therapy and ankle pump exercise [ OR=3.013, 95% CI (0.116-0.789)] was statistically significant (P<0.05) and was a high risk factor for deep vein thrombosis. CONCLUSION: The incidence of DVT is still high in patients with spinal cord injury after surgery. When one or more risk factors such as age >45 years, history of diabetes, smoking history, and ASIA grade A of spinal cord injury is present, preventive measures should be taken in advance, it is of great significance to prevent the formation of deep vein thrombosis.

A biopsychosocial model of resilience for breast cancer: A preliminary study in mainland China.

PURPOSE: Patients diagnosed with breast cancer exhibited critical biopsychosocial functions following surgery or adjuvant treatment; therefore, it is important that they exhibit resilience. A Resilience Model for Breast Cancer (RM-BC) was developed using Chinese breast cancer patients to increase our understanding of how resilience outcomes are positively and negatively affected by protective and risk factors, respectively. METHODS: Chinese women with breast cancer completed the questionnaires within 1 week of beginning treatment. Exploratory Structural Equation Modeling was used to evaluate the RM-BC using a sample size of 342 patients. RESULTS: RM-BC suggested satisfactory goodness-of-fit indices and 67 percents of variance for resilience was explained. The Fit Indices for the measurement model were as follows: CFI = 0.909, GFI = 0.911, IFI = 0.897, NFI = 0.922, PNFI = 0.896, PCFI = 0.884, and RMSEA = 0.031. Three risk factors - emotional distress, physical distress, and intrusive thoughts - and four protective factors - self-efficacy, social support, courage-related strategy, and hope - were recognized. CONCLUSION: The resilience model allows for a better understanding of Chinese breast cancer patients' resilience integration while undergoing treatment and provides an effective structure for the development of resilience-focused interventions that are grounded in their experiences. A randomized trial has provided evidences of feasibility in Chinese women with breast cancer and the resilience model could be used as a useful framework for more resilience intervention in the future.

Psychometric properties of the Chinese version of resilience scale specific to cancer: an item response theory analysis.

OBJECTIVE: Classic theory test has been used to develop and validate the 25-item Resilience Scale Specific to Cancer (RS-SC) in Chinese patients with cancer. This study was designed to provide additional information about the discriminative value of the individual items tested with an item response theory analysis. METHODS: A two-parameter graded response model was performed to examine whether any of the items of the RS-SC exhibited problems with the ordering and steps of thresholds, as well as the ability of items to discriminate patients with different resilience levels using item characteristic curves. RESULTS: A sample of 214 Chinese patients with cancer diagnosis was analyzed. The established three-dimension structure of the RS-SC was confirmed. Several items showed problematic thresholds or discrimination ability and require further revision. CONCLUSIONS: Some problematic items should be refined and a short-form of RS-SC maybe feasible in clinical settings in order to reduce burden on patients. However, the generalizability of these findings warrants further investigations.

The role of RANKL/RANK/OPG system in the canine model of hip periprosthetic infection osteolysis.

PURPOSE: We aimed to investigate whether the RANKL/RANK/OPG system is associated with the incidence of periprosthetic osteolysis with septic loosening, and to investigate the differences of RANKL/RANK/OPG system expression in synovial fluid surrounding the normal and septic loosening hip prosthesis in canine models. METHODS: Twelve healthy adult mongrel canines were divided into two groups: experimental and control. Femoral head and stem replacements were conducted on the right side in both groups. The experimental group received the bacteria fluid intra-articular injection and the other group received the same amount of saline in the same day. The synovial fluid samples were gathered at the 1st, 2nd, 4th, 8th, 12th, 16th and 19th week after the bacteria fluid intra-articular injection for enzyme-linked immunosorbent assay (ELISA), the expression of the RANKL/RANK/OPG system. RESULTS: Surgery on all animals was successful. Two dogs were excluded from the analysis of the result because of a surgery infection or death. The ELISA of the synovial fluid revealed that the ratio of RANKL/OPG showed a significant upward trend (p≤0.05) with time in the test group but the ratio of RANKL/OPG in the control group changed slowly over time (p>0.05). The ratio of RANKL/OPG value between the test and control group showed a significant upward trend, but had no statistical difference (p>0.05) over time. CONCLUSIONS: It could be concluded that the RANKL/RANK/OPG system is correlated with the incidence of periprosthetic osteolysis with septic loosening. Consequently, imbalance RANKL/RANK/OPG system was related to periprosthetic osteolysis with septic loosening.

[Establishment of arthroscopic trans-septal approach and its clinical application].

OBJECTIVE: To investigate the method and result of arthroscopic trans-septal approach (ATS). METHODS: Ten fresh cadaveric knees were prepared for anatomical study about the posterior septum, and 65 posterior compartment arthroscopy of the knees were performed to view the structure of the posterior septum. The initial diagnosis included: rheumatoid arthritis, pigmented villonodular synovitis, osteoarthritis, loose body or foreign body in the posterior compartment, posterior cruciate ligament (PCL) injury or avulsion fracture, posterior horn tear of meniscus, undiagnosed swollen knee with pain and effusion, osteochondritis dissecans, pyogenic arthritis, gout. From January 2002 to June 2005, 22 cases of ATS were applied. Anterolateral portal was initially created, followed by posterolateral portal under the viewing of arthroscopy which was located at the anterolateral portal. Anteromedial and posteromedial portals were also created using the same technique. Arthroscopy was then transferred to the posteromedial portal, and blade was introduced from the anteromedial portal to gradually remove the synovium covering PCL. Arthroscopy was relocated to the anteromedial portal, Wissinger rod was introduced from the posteromedial portal and pointed to the posterior septum adjacent to the posterior edge of the midportion of PCL. The Wissinger rod was pushed carefully to pierce through the posterior septum under the sight of arthroscopy which was located at the posterolateral portal. ATS was finally created. RESULTS: The posterior septum was in the middle of posterior compartment of the knee, which was film screen-like at the sagittal plane and sandwich-like at the transverse plane. The synovium covered the posterior septum at arthroscopic inspection. Twenty-two cases of ATS were successfully created, amounting to 34% (22/65) of all cases at the same period which had received the arthroscopy of posterior compartments of the knees. Synovectomy of the posterior compartments of the knees was performed in 7 cases, loose body removal was in 6 cases, PCL reconstruction was in 4 cases, reduction and fixation of PCL avulsion fracture was in 2 cases. Chondroplasty, inflammatory synovectomy, and meniscectomy were performed accordingly in 6 osteoarthritis cases. No vascular or nervous injury was encountered. At an average of 20 months follow-up (range, 4 to 45 months), 9 cases still had mild knee pain or swelling, 2 cases had severe pain and were recommended for total knee replacement, the other 11 cases had no recurrence of knee pain or swelling. CONCLUSIONS: ATS has no blind area under arthroscopic vision and facilitate trans-septal operation. It is a safe and effective method to treat the diseases of the posterior compartment of the knee. The direction of inside to outside to create ATS is comparatively reliable, and PCL could be identified as an interior landmark during the passage of Wissinger rod through posterior septum to create ATS.

[Study on the patterns of TCM syndrome differentiation of 900 patients with posthepatitic cirrhosis].

OBJECTIVE: To investigate the formula for TCM syndrome pattern differentiation in patients with posthepatitic cirrhosis. METHODS: Factors analysis, C mean clustering and fuzzy comprehensive assessment were used for analyzing the basic information, including clinical symptoms and signs, obtained from 900 patients with posthepatitic cirrhosis to establish the formula for syndrome pattern differentiation through collecting the syndromic factors. RESULTS: Multivariate statistic analysis showed the syndromic factors could be classified into two categories: the common characteristics related factors, which reflected the basic pathology of cirrhosis as qi deficiency and blood stasis, and the factors for differentiating 5 syndrome patterns (Gan-Shen yin-deficiency, dampness-heat accumulation, stasis-heat accumulation, Gan-qi stagnation with Pi-deficiency, and Pi-Shen qi-deficiency), which reflected the different pathological features of different syndrome patterns, showing the complexity and polymorphism of syndrome construction. And the functional formula for differentiating syndrome pattern was established. CONCLUSION: The differentiating formula of TCM syndrome pattern could provide a quantified criterion for syndrome pattern differentiation in posthepatitic cirrhosis patients.